LOCUS       Exported                4845 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046315
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4845)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4845)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4845
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             514..531
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             532..555
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(600..618)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             969..1772
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      1855..2837
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2915..2944
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2966..3553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3565..3592
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3616..3807)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3808..4668)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4431..4450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4669..4771)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      4772..4803
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gctgatgcac aatcgttttt aaacgggttt gcgctcgtgc cgagaggttc ttggagccat
      541 ccgcagtttg aaaaataaac aggccccttt tcctttgtcg atatcatgta attagttatg
      601 tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga
      661 caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt
      721 tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc
      781 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
      841 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
      901 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
      961 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     1021 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     1081 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     1141 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     1201 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     1261 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     1321 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     1381 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     1441 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     1501 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     1561 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     1621 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     1681 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     1741 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     1801 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa
     1861 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     1921 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     1981 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     2041 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     2101 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     2161 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     2221 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     2281 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     2341 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     2401 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     2461 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     2521 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     2581 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     2641 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     2701 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     2761 gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa
     2821 gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg ttacaagctt
     2881 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc
     2941 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3001 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3061 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3121 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3181 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3241 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3301 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3361 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3421 cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc tttatagtcc
     3481 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg
     3541 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt
     3601 acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3661 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3721 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3781 aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg cacctatctc
     3841 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3901 gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3961 accggctcca gatttatcag caataaacca gccagccggg agggccgagc gcagaagtga
     4021 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4081 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4141 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4201 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgccag
     4261 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4321 tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4381 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4441 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact
     4501 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4561 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4621 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt tagcttcctt
     4681 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4741 gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg
     4801 actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
